3,21-disubstituted-5,7,9 (11)-pregnatrien-20-ones



United States Patent 2,813,106 3,21-DISUBSTlTUTED-5,7,9 (11)-PREGNATRIEN- 20-ONES Robert H. Levin, A Vern McIntosh, Jr., and George B.

Spero, Kalamazoo, Mich, assignors to The Upjohn Company, Kalamazoo, Mich., a corporation of Michigan No Drawing. Application September 1, 1950, Serial No. 182,882

Claims. (Cl. 260397.45)

1 The present invention relates to 3,2l-di-substituted- 5,7,9(1l)-pregnatrien-20-ones, and to a process for the production thereof. The present application is a continuation-in-part of our prior filed copending application Serial No. 136,656, filed January 3, 1950.

The compounds of the present invention may be represented by the structural formula:

AcO l wherein X is selected from the group consisting of hydrogen and acetyl, and wherein Ac is selected from the group consisting of hydrogen and the residue of an organic carboxylic acid containing from one to eight carbon atoms, inclusive.

It is an object of the present invention to provide a novel group of compounds which are useful in the preparation of steroid compounds containing an oxygen atom at carbon atom eleven. Another object of the invention is the provision of a process for the production of novel compounds, the 3,2l-disubstituted-5,7,9(1l)-pregnatrien 20-ones of the above formula. Other objects of the invention will become apparent hereinafter.

The compounds of the present invention, as previously stated, are useful in the preparation of steroid compounds having an oxygen atom attached to carbon atom eleven. Such compounds are of particular interest in the field of steroid research due to the biological activity of the cortical hormones and certain known derivatives thereof, which oxygenated steroids are known to have biological effects differing markedly from the unoxygenated steroids. It is, therefore, of importance to investigate the oxygenated derivatives of such steroids, particularly those oxygenated at carbon atom eleven, as well as to investigate the biological activity of the adducts themselves and their transformation products. The importance of such investigation is moreover emphasized by the acute shortage of adrenal cortical hormones, and the absence of any present suggestion for alleviation of the said shortage except through organic synthesis.

Novel compounds of the present invention which are of particular interest are those compounds of the above generic formula wherein AcO represents an ester of the 3-hydroxy group with a carboxylic acid containing up to and including eight carbon atoms. Among the acids -which can be used are formic, acetic, propionic, butyric,

valeric, hexanoic, heptanoic, octanoic, succinic, glutaric, cyclopentanoic, cyclohexanoic, benzoic, toluic, naphthoic, and the like. Preferred acids are the lower-aliphatic acids. The acids may also contain substituents, such as halo, alkyl, and methoxy, which are non-reactive under the reaction conditions employed.

The compounds of the present invention are prepared by removal of the maleic acid or maleic anhydride radical from a 3,21-disubstituted-5,7,9(11)-pregnatrien-20-one of the formula:

wherein X is selected from the gen and acetyl, wherein Ac is selected from the group consisting of hydrogen and the residue of an organic carboxylic acid, all as previously defined, and wherein A is the adduct radical of maleic acid or maleic. anhydride.

Such adducts, wherein X is acetyl, are conveniently prepared by reaction of the selected 3-acyloxy-5,7,9( l l)-pregnatrien-ZO-one or 3hydroxy-5,7,9 l l -pregnatrien-20-one maleic acid or anhydride adduct with lead tetraacetate. The lead tetraacetate is ordinarily employed in an amount in excess of the theoretical, and an organic solvent for the reaction is employed, for example acetic acid and water, benzene, or ether, with acetic acid being preferred. The reaction may conveniently be carried out at a temperature between about twenty and degrees centigrade, with temperatures of about 50-60 degrees centigrade, which are readily obtainable by employment of a waterbath, being preferred. The lead tetraacetate usually goes into solution quite readily, and the reaction is complete in from 24 to 96 hours. The Zl-acetoxy adduct may then be recovered by evaporating the solution under a vacuum, dissolving the residue in ether, washing with .water, drying, and recrystallizing from a solvent, such as alcohol, if desired, or in other conventional manner. The 3-acyloxy '21-hydroxy and 3,2l-dihydroxy adducts are prepared by C: O H-Acyloxy AcO wherein A and Ac have the values previously given. v

Adducts of 3,22 diacyloxybisnor 5,7,9( 1 l),20 (22)- cholatetraenes [22-enol esters of 3-acyloxybisnors- 5,7,9(11) cholatrien-22-als] are conveniently prepared by I CHCHO CH: i

wherein A and. Ac have the values previously given, to the action of an acid anhydride'or an acid halide in the presence of an alkaline salt of the acid. The starting adducts of 3-acyloxybisnor-5,7,9( l1)-cholatrien-22-als can be prepared from adducts of 3-esters of dehydroergosterol by selective oxidation as described and claimed in the copending application Serial No. 111,100 of Robert H. Levin, filed August 18, 1 949, and as more fully described hereinafter.

The 3-esters of dehydroergosterol, from which the 3- acyloxybisnor-5,7,9(l 1) cholatrien-22-al adducts are prepared, can be synthesized in several ways starting with ergosterol. For example, erogsterol can be transformed to dehydroergosterol with mercuric acetate according to known methods [Windauset al., Ann. 465, 157 (1928)] and the I i-hydroxy group of the dehydroergosterol acylated by knownprocedure. Alternatively the 3-hydroxy group of ergosterol, can] be acylated prior to the preparation of the dehydrfo derivative, a procedure which is particularly preferred in the preparation of the 3-acetoxy derivative. Thefadducts of dehydroergosterol are then prepared by the addition of maleic anhydride or the like to dehydroer'gosterolor a3-ester thereof according to known methods [Honigmann, Ann. 508, 89 (1934)]. The anhydrides can then be converted to their corresponding acids and esters if desired.

The ester group, when present in the 3-position of dehydroergosterol, is for the purpose of protecting the 3- hydroxy group in subsequent chemical reactions. For

this purpose any convenient ester of an organic carboxylic acid. which is non-reactive under the conditions of the reaction, is snitablle. The preferred acids are the fatty acids such. as formic, acetic, propionic, butyric, valeric, hexanoic, heptanoic, octanoic; dibasicacids such as malonic, succinic, phthalic; cycloaliphatic acids such as cyclopentanoic and cyclohexanoic; and aromatic acids such as benzoic, toluic, and the like. The acids may also contain substituen ts such as halogen, alkyl, the methoxy radical, and the like, and these substituents will be carried throughout: the synthesis. Ifdesired, the acyl group can be changed to another acyl. group by saponifying the ester to give a 3-hydroxy compound, which can then be reesterified as previously described.

Apreferredrnethod for preparing some of the dehydroergosteryl adducts comprises the saponification of a 3- acyloxy-adduct of dehydroergosterol with dilute alkali followed by acidification. The 3-hydroxy di-carboxylic acid thus formed can be converted to the S-hydroxy anhydride by heat, or it can be converted to any desired 3- acyloxy anhydride adduct by heating under reflux with the appropriate acid anhydride or chloride in pyridine solution. Dialkyl esters of the previously mentioned dicarboxylic acid adducts can be prepared by subjecting the acid to the action of an esterification reagent such as diazoalkanes [Wilds et al., J. Org. Chem. 13, 763 (1948)] e. g., diazomethane, diazoethane, diazohutane, and the like. i

The selective oxidation of an adduct of dehydroergosterol, or a 3-ester thereof, to produce an adduct of 3-hydroxy-bisnor-5,7,9(11)-cholatrien 22:al, or a 3-ester thereof, is. accomplished by dissolving the, dehydroergosteryl adduct in a suitable solvent, cooling to about minus to plus 30 degrees centigrade, and passing ozone into the solution until about 1.0 to 1.25 moles of ozone per mole of adduct have been absorbed. The temperature of the solution should be maintained below plus 30 degrees centigrade, preferably between a temperature of minus 30 and minus 70 degrees centigrade, during the addition of ozone, although temperatures as low as minus 80 and as high as plus 30 degrees centigrade are operative. The lower temperatures of the preferred range are readily obtained by cooling the solution of the adduct with a bath of solid carbon dioxide in acetone or the like, although various other methods of cooling can be used. Many of the customary solvents used in ozonizations such as chloroform, acetic acid, carbon tetrachloride, ethylene chloride, methylene chloride, and the like, can be used.

The ozonides are then decomposed under reducing conditions, that is, in the absence of oxidizing agents, whether added or formed in the course of the reaction by products of decomposition of the ozonide. This means that excess oxygen formed by decomposition of the ozonide is prevented from forming hydrogen peroxide by combining with any moisture present, and that molecular oxygen is prevented from oxidizing the aldehyde thus formed. This can be conveniently accomplished by decomposing thc ozonide in glacial acetic acid by the addition of finelypowdered zinc.

As is conventional with ozonizations when conducted in solvents, other than glacial acetic acid, the solvent used for ozonization is replaced, after completion of the ozonization, by adding glacial acetic acid and removing the lower-boiling solvent by fractional distillation. Alternatively, the solvent can be removed by careful warming under reduced pressure prior to the addition of glacial acetic acid, if desired.

After decomposition of the ozonide and removal of the zinc, the aldehyde can be recovered by diluting the acetic acid with water, or in other conventional manner, such as by formation of an aldehyde derivative, e. g., the dinitrophenylhydrazone.

Adducts of 3,22-diacyloxybisnor-5,7,9(1l),20(22) cholatetraenes [22-enol-esters of adducts of S-acyloxybisnor-5,7,9(ll)-cholatrien-22-als] can be conveniently, prepared by heating the corresponding 3-hydroxy or 3- acyloxy aldehyde maleic acid, maleic acid anhydride, or maleic acid ester adduct with a large excess of an organic carboxylic acid anhydride in the presence of a small amount of the alkali metal salt of the acid corresponding to the anhydride employed or an acid catalyst such as para-toluene sulfonic or sulfuric acid, The preferred anhydride is acetic anhydride, but other anhydrides, such as propionic, butyric, valeric, hexanoic, and octanoic anhydrides, as well as benzoic acid anhydride, ortho-toluic acid anhydride, and the like, are also operative. The acid anhydrides can also be substituted by non-reactive groups, such as halo, alkyl, and methoxy, as in the case of;chlor o.- acetic, ortho-toluic, or methoxybenzoic acid anhydrides. The reaction can be conveniently followed by observing the color changes in the reaction mixture, optimum yields being obtained 'by discontinuing the application of heat when the color of the solutionchanges from yellow to brown. Ordinarily the reaction is heated at about degrees centigrade for from about four to six hours, but temperatures as low as 100 and as, high as degrees centigrade are also operative. Thereaction is usually conducted at;the'boiling point of the anhydride, but in the case of the higher-boiling anhydrides, such as benzoic anhydride, a suitable temperature control, such as 100-150 degrees centigrade, must be used, since the adduct otherwise tends to decompose, in the higher temperature range. If a 3-hydroxy aldehyde adduct is thus reacted with an anhydride, the hydroxy groupwill be acylated, and, similarly, if a 'maleic acidzadduct is usediinstead of a diester or an anhydride, the anhydride will be formed. The enol ester can be 'isolatedby removing the excess anhydride 'ductive conditions phenylhydrazone.

.imder reduced pressure, and separating the enol ester from alkali metal salts, which procedure yields a product sufficiently pure for most purposes, but which can be further purified by recrystallization from acetone-water, acetonepentane, or like solvents, if desired.

The ozonization of the thus-prepared enol acylate to produce 3-acyloxy-5,7,9(11)-pregnatrien-20-one adducts involves dissolving the enol ester in a suitable solvent, cooling to about minus eighty degrees c'entigrade to plus thirty degrees centigrade, and passing ozone, ozonized air, or ozonized oxygen into the solution until about 1.0 to about 1.25 moles, preferably 1.0 to 1.1 moles, of ozone per mole of adduct has been absorbed. The addition of ozone to the 20:22 double bond is so rapid that only 'a small amount of ozone escapes from the reaction mixture, and the amount of ozone ordinarily required there- 'fore closely approximates the theoretical amount. Loss to the solvent, if any loss occurs, must be taken into consideration in calculating the amount of ozone to be introduced. The temperature of thesolution should be maintained below plus thirty degrees centigrade, preferably between a temperature of minus thirty and minus seventy degrees centigrade, during the addition of ozone, although temperatures as low as minus eighty and as high as plus thirty degrees centigrade are operative. The

lower temperatures of the range are readily obtained by cooling the solution of the adduct with a bath of solid carbon dioxide in acetone or the like, although various other methods of cooling may be employed. Many of the customary solvents used in ozonizations, such as chloroform, methylene 'chloride, ethylene chloride, carbon tetrachloride, acetic acid, and the like, can be used for the ozonization reaction.

The 20:22 ozonides thus produced are then decomposed under conditions normally employed for decomposition of such compounds. This can conveniently be accomplished by decomposing the ozonide with hydrogen peroxide, by hydrolysis, by treatment with zinc in glacial acetic acid, or by a catalytic amount of colloidal metal such as silver, platinum, or palladium in a solvent, such as glacial acetic acid, alcohol, or ethyl acetate, in which latter case reductive conditions, e. g., a hydrogen atmosphere are also employed. The use of reis well established in the art [Hill and Kelly, Organic Chemistry, page 63, Blackiston Company, Philadelphia (1943); Church et al., I. Am. Chem. Soc. 56, 176484 (1934); Gilman, Organic Chemistry," second edition, page 63, John Wiley and Sons, New York 1943) Long, Chem. Reviews 27, 452454 (1940)].

As is conventional with decomposition of ozonides with zinc, when the ozonizations are conducted in solvents other than glacial acetic acid, the solvent used for'the ozonization is replaced, after completion of the ozonization, by adding glacial acetic .acid and removing the lower-boiling solvent by fractional distillation, or the solvent can be removed by careful warming under reduced pressure prior to the addition of acetic acid, if

desired. After decomposition of the 20:22 ozonide and removal of the metal, the ketone can be recovered by diluting the acetic acid with water, or by other conventional procedure for the recovery of ketones, such as by formation of a carbonyl derivative, e. g., the 2,4-dinitro- Recrystallization from acetone or the like results in a more highly purified ketone product.

The method of the invention consists in heating a 3,21- disubstituted-5,7,9(11)-pregnatrien-20-one maleic acid or anhydride adduct in the presence of an organic amine ata temperature of about 100 to 225 degrees centigrade, "with or without the presence of an inert organic solvent, and thereafter isolating the product. The method of the invention has the advantage of being conveniently applicable to large scale work in that it is not necessary to remove the components from the reaction mixture to complete the reaction. I t'also has the further advantage r v-cvfa'r 1., t ,t..

6 that the desired triene can be obtained in a high degree of purity and in excellent yields.

Amines that can be used in the process of the invention are: secondary aliphatic amines such as dimethylamine, diethylamine, dipropylamine, dibutylamine, diamylamine, dioctylamine; tertiary aliphatic amines such as trimethylamine, triethylamine, triamylamine, methyldioctylamine, methyldiethylamine; secondary and tertiary cycloaliphatic amines such as N-methylcyclohexylamine, N,N-dimethylcyclohexylamine, N,N-diethylcyclohexylamine; secondary and tertiary heterocyclic amines such as pyrrolidine, N-methylmorpholine, N-ethylpyrrolidine, morpholine, piperidiue, N-methylpiperidine, Z-methylpiperidine, 1,2-dimethylpiperidine, 1,2,4-trimethylpiperidine, 2,4,6-trimethylpiperidine, 1-ethyl-2,4,6-trimethylpiperidine; aromatic heterocyclic amines such as pyridine, picoline, lutidine, collidine, quinoline, quinaldine, lepidine, 3-methylquinoline; secondary and tertiary carbocyclic aromatic amines, such as N-methylaniline, N-ethylaniline, N-butylaniline, N-benzylaniline, N,N-dimethylaniline, N,N-diethylaniline, N,N-dibutylaniline, N,N-dibenzylaniline, N-methyltoluidine, N,N-diethyltoluidine, N-ethylxylidine, N,N-dimethylxylidine; substituted aliphatic amines such as diethylaminoethanol, dibutylaminoethanol, N-pyrrolidylethanol, N-piperidylethanol; substituted aromatic amines such as orthomethoxy-N,N-dimethylaniline, para-cthoxy-N,N-diethylaniline, parachloro-N,N- dimethylaniline, para-bromo-N,N-diethylaniline, parafluoro-N,N-dibutylani1ine, N,N-dimethylmesidine; secondary and tertiary aralkyl amines such as methylbenzylamine, dimethylbenzylamine, propylbenzylamine, diisopropylphenethylamine, diethylphenylisopropylamine; and primary amines such as butylamine, hexylamine, octylamine, cyclohexylamine, aniline, toluidine, xylidine and the like.

The process of the present invention, then, comprises heating a selected 3,21-disubstituted-5,7,9(11)-pregnatrien-20-one maleic acid or anhydride adduct, of the formula given previously, to a temperature between about and 225 degrees centigrade, preferably between about and 200 degrees centigrade, in the presence of an organic amine, removing excess amine, and recovering the product, wherein the maleic acid or maleic anhydride adduct has been eliminated from the molecule, with restoration of the conjugated double bond system at carbon atoms 5,6:7,8. The time required for the reaction is usually from about one to eight hours, depending upon variable factors such as the particular adduct treated, the amine employed, and the temperature of reaction. Ordinarily, a reaction period of about four hours, is entirely satisfactory, although, at the lower temperatures, a more extended period may be employed to advantage. The employment of pressure may in some cases be advantageous although it is in most cases preferred to conduct the pyrolysis reaction at atmospheric pressure.

The following examples are illustrative of the process and products of the present invention, but are not to be construed as limiting.

PREPARATION 1.-DIMETHYL MALEATE ADDUCT 0F Derry- DROERGOSTERYL BENZOATE To a solution of 21 grams of dimethyl maleate adduct of dehydroergosterol in 69 milliliters of warm pyridine was added 9.5 milliliters of benz-oyl chloride. After standing at room temperature for fifteen minutes, the mixture was poured into 1400 milliliters of ice-water and the solid removed by filtration, dried, and recrystallized from acetone. There was thus obtained 26.4 grams of dimethyl maleate adduct of dehydroergosteryl benzoate, melting at 203 to 205.5 degrees centigrade.

PREPARATION 2.DIMETHYL MALEATE Anuucr OF DEHY- DROERGOSTERYL ACETATE In a manner essentially that described in Preparation A 1, the dimethyl maleate adduct of dehydroergosteryl acetate, melting at 177 to 179 degrees centigrade, was prepared from the dimethyl maleate adduct of dehydroergosterol and acetyl chloride.

P PARAT ON 3.-DIMETHYL MALEATE Annucr F DEHY- D oERoosTERYL FoRMATE A solution of six grams of dimethyl maleate adduct of dehydroergosterol in fifty milliliters of 87 percent formic acid was heated under refiux for one hour, cooled, and the dimethyl maleate adduct of dehydroergosteryl formate filtered therefrom. Upon crystallization from aceetone, the purified material melted at 177.5 to 178.5 degrees centigrade.

PREPARATION 4.MALEIC Acm ADDUCT OF DEl-IYDRO- ERGOSTEROL A solution of 2.0 grams of sodium hydroxide in twenty milliliters of water was added to a solution of 1.73 grams of the maleic anhydride adduct of dehydroergosteryl acetate (M. P. 230-232 degrees centigrade) in forty milliliters of dioxane. The mixture solidified but dissolved on addition of 300 milliliters of water and heating to eighty degrees centigrade. After half an hour the solution was cooled and made acid with aqueous three normal hydrochloric acid, to give 1.61 grams of precipitate. On crystallization from a dioxane-water mixture, the maleic acid adduct of dehydroergosterol melted at 190-192 degrees centigrade.

PREPARATION 5.-MALEIc ANHYDRIDE ADDUCT 0F 3-HEP- TANOYLOXYDEHYDROERGOSTEROL The maleic acid adduct of dehydroergosterol from Preparation 4 was dissolved in a mixture of seven milliliters of warm pyridine and fourteen milliliters of heptylic anhydride, and the mixture heated under reflux for one hour. About eighty percent of the reaction solvent was removed under reduced pressure, and the residue then dissolved in methyl alcohol. The methyl alcohol solution was concentrated and cooled to yield 4.8 grams of the maleic anhydride adduct of 3-heptanoyloxydehydroergosterol, melting at 186-1915 degrees centigrade.

PREPARATION 6.MALEIC ANHYDRIDE ADDUCT 0F B-BETA- AcEToxYBIsNoR-5 ,7 ,9 1 1 -cIIoLArRIEN-22-AL A solution of 5.35 grams of the maleic anhydride adduct of 3 beta acetoxydehydroergosterol in 107 milliliters of methylene chloride was cooled to about minus seventy degrees centigrade and ozonized until 505 milligrams of ozone had been absorbed. The temperature of the solution was then gradually raised to about plus ten to fifteen degrees centigrade, whereupon seventy rnilliliters of glacial acetic acid was added and the methylene chloride removed under reduced pressure. Seven grams of zinc dust was then added to the cold solution at a uniform rate over a period of ten minutes, While keeping the reaction temperature below plus twenty degrees centigrade. After beingstirred for fifteen minutes, the mixture was filtered and the filtrate poured into water. There was thus obtained 4.31 grams of maleic anhydride adduct of 3 beta acetoxybisnor 5,7,9(11) ch-olatrien 22- al, a fine whiteipowder which melted at 187-197 degrees centigrade.

' To a solution of 0.30 gram of the maleic anhydride adduct of.3 -.beta acetoxybisnor 5,7,9(11) cholatrien 22 al, in thirty milliliters of ethanol, was added twenty millilters of alcohol containing one percent 2,4- dinitrophenylhydrazine and three percent concentrated hydrochloric acid. The mixture was allowed to stand for one hour at room temperature and then placed in a refrigerator to complete precipitation of the yellow crystals. The precipitate was then collected and recrystallized from a mixture of chloroform and alcohol, to give the 2,4- 'dinitrophenylhydrazone of the maleic anhydride adduct of 3 beta -acetoxybisnor 5,7,9 (11) cholatrien 2 2 al,

:meltingat 269-271degrees-centigrade.

PREPARATION 7 .-MALEIc ANiIYDRmE Ammo-r 0F 3 min- AcaToxYBIsNoR-5,7,9(11)-cIIoI.ArRIEN-22-AL A two-liter, round-bottom flask was charged with fifty grams (0.93 mole) of dehydroergosteryl acetate maleic anhydride adduct and one liter of methylene chloride. The solution was cooled to Dry-Ice temperature with a trichloroethylene bath and ozonized oxygen passed at a rate of 1200 milliliters of oxygen per minute (at this rate the ozonizer was producing about 36 milligrams of ozone per minute). The flow of ozonized oxygen was maintained for 128 minutes, a total of 4608 milligrams percent) of ozone being passed into the solution. The reaction mixture was transferred to a two-liter, round-bottom flask fitted with a capillary and a com denser for downward distillation, 300 milliliters of acetic acid added, and the methylene chloride distilled over in vacuo at forty degrees centigrade or below. The flask was then placed in a water bath and fitted with a stirrer. An additional 200 milliliters of acetic acid was added and the ozonide decomposed by the addition of fifty grams of zinc dust. The zinc dust was added in portions over a period of twenty to thirty minutes while the solution was stirred and the temperature maintained at seventeen to twenty degrees centigrade. After addition, the mixture was stirred for another twenty minutes and then filtered. The precipitated zinc dust was washed by filtering 100 milliliters of acetic acid therethrough, and the filtrate gradually diluted with water (1100 to 1200 milliliters) until the product had been drowned out. The product was then cooled in the refrigerator overnight and filtered. The yield of crystalline product was 42 grams, assaying 89-95 percent of the desired aldehyde.

PREPARATION 8 In a manner essentially that described in Preparation 6, the following compounds were prepared.

(1) Maleic anhydride adduct of 3-beta-formoxybisnor- 5,7,9(1l)-cholatrien-22-al, melting at 95-130 degrees centigrade. 2,4-dinitrophenylhydrazone, melting at -168 degrees centigrade.

(2) Maleic anhydride adduct of 3-betal-heptanoyloxybisnor-5,7,9( ll)-cholatrien-22-al, melting at 197.5-199 degrees centigrade. 2,4-dinitrophenylhydrazone, melting at 253-257 degrees centigrade.

(3) Dimethyl maleate adduct of 3-beta-benzoyloxybisnor-5,7,9(1l)-chloratrien-22-al, melting at 183-187 degrees centigrade. 2,4-dinitrophenylhydrazone, melting at 224-249 degrees centigrade.

(4) Dimethyl maleate adduct of 3-beta-acetoxybisnor- 5,7,9(ll)-cholatrien-22-al, melting at 172-178 degrees centigrade. 2,4-dinitrophenylhydrazone, melting at 238 to 244 degrees centigrade.

(5) Dimethyl maleate adduct of 3-hydroxybisnor-5,7,9- (ll)-cholatrien-22-al, melting at 163-170 degrees centigrade. 2,4-dinitrophenylhydrazone, melting at 250-254 degrees centigrade.

In a manner similar to the above, the maleic anhydride adduct of 3-hydroxybisnor-5,7,9( l1)-cholatrien-22-al is obtained from dehydroergosteryl maleic anhydride adduct; the maleic acid adduct of 3-hydroxybisnor-5,7,9( 11)- cholatrien-22-al is obtained from dehydroegosteryl maleic acid adduct; and 3-acyloxybisnor-5,7,9(11)-cholatrien-22-al maleic acid adducts are obtained from the maleic acid adduct of 3-acyloxy-dehydroergosterols.

PREPARATION 9.DIMETHYL MALEATE Annucr 0F 3-HY- DROXYBISNOR-5 ,7 ,9( 1 1 )-crroLATRIEN-22-AL A solution of 2.69 grams (.005 mole) of the dimethyl ester of the maleic acid adduct of dehydroergosterol, in eighty milliliters of methylene chloride, cooled by a Dry- Ice and trichloroethylene bath, was treated with ozonized oxygen until 247.36 milligrams (.0051 mole) of ozone was absorbed. The solution was then allowed to warm to room temperature, whereafter thirty milliliters of acetic acid was added and the methylene chloride removed in vacuo.

While cooling in a water-bath at fifteen degrees centigrade, four grams of zinc dust was added in portions with stirring, the temperature being maintained between fifteen and twenty degrees centigrade. Stirring was continued for another fifteen minutes, whereafter the zinc was separated by filtration. The filtrate was diluted with water to cloudiness, extracted with ether, the ether extract washed with sodium bicarbonate and then with water to neutrality, the solution then dried over sodium sulfate and evaporated to dryness in vacuo. The residue was crystallized from acetic acid and water, giving 1.92 grams (81.5 percent of the theoretical), melting point 91-97 degrees centigrade, which yielded a dinitrophenylhydrazone derivative in 72.5 percent yield, melting point 212-238 degrees centigrade. The aldehyde was recrystallized and found to have a purified melting point of 163-170 degrees centigrade, while the dinitrophenylhydrazone derivative was recrystallized until a melting point of 250-254 degrees centigrade was attained.

PREPARATION 10.MALEIC ANHYDRIDE ADDUCT OF 3-BETA- ncaroxr 22 ACETOXYBISNOR 5,7,9(11),20(22)-cno- LATETRAENE A mixture of twenty grams of the maleic anhydride adduct of 3-beta-acetoxybisnor-5,7,9(11)-ch0latrien-22-al, six grams of anhydrous sodium acetate, and 600 milliliters of acetic anhydride, was heated under reflux for six hours, whereafter volatile components were removed under reduced pressure. The resulting solid was digested with five fifty-milliliter portions of boiling acetone for five minutes each, and the extracts combined and diluted with 130 milliliters of water. There was thus obtained sixteen grams of the maleic anhydride adduct of 3-beta-acetoxy-22-a-cetoxybisnor-5,7,9(1l),20(22)-cholatetraene, which melted at 186 to 193 degrees centigrade. Recrystallization of the crude product from a mixture of acetone and pentane raised the melting point to 200.5 to 202 degrees centigrade.

PREPARATION 11 In a manner essentially that described in Preparation 10, the following compounds were prepared:

(1) The dimethyl maleate adduct of 3-beta-benzoyloxy- 22-acetoxybisnor-5,7,9( 11),20(22)-ch0latetraene, which melted at 210 to 211 degrees centigrade.

(2) The dimethyl maleate adduct of 3-beta-acetoxy-22- acetoxybisnor-5,7,9 1 1 ,20 (22) -cholatetraene, which melted at 181 to 183 degrees centigrade.

In the same manner as given above, 22-acyloxy, e. g. formoxy, acetoxy, propionoxy, butyroxy, valeroxy, hexanoyloxy, heptanoyloxy, octanoyloxy, benzoloxy, naphthoyloxy, and the like 3-acyloxybisnor-5,7,9(l1),20(22)- ch-olatetraene adducts, are obtained from the compounds of Preparations 6, 7 and 8. Such representative compounds include 3-formoxy-22-acetoxybisnor-5,7,9(11),20- (22)-cholatetraene, 3-propionoxy-22-acetoxybisnor-5,7,9- (11),20(22) cholatetraene, 3,22-dipropionoxybisnor-5,7, 9 1 1 -20(22) -cholatetraene, 3,22-dibenzoyloxybisnor- ,7 9( 1 1) ,20 (22) -cholatetraene, and 3-heptanoyloxy-22-octaoyloxybisnor-S ,7,9( 11),20(22)-cholatetraene adducts with maleic anhydride or maleic acid esters such as the dimethyl maleate, diethyl maleate, dipropyl maleate, diisopropyl maleate, dibutyl maleate, dioctyl maleate, dibenzyl maleate and the like.

PREPARATION l2.-MALEIC ANHYDRIDE ADDUCT OF 3-BETA- ACETOXY-5 ,7 ,9( 1 1 -PREGNATRIBN-20-ONE the maleic anhydride adduct of 3-beta-acetoxy-5,7,9(11)- pregnatrien-ZO-one, which melted at 240 to 264.5 degrees centigrade. Several recrystallizations of the crude material from acetone raised the melting point to 263.5 to 264.5 degrees centigrade.

PREPARATION 13.-MALBIC Acn) AnnUcroF 3-BE'IA- HYDROXY-5,7,9 1 1 -PREGNA'IRIEN-20-ONE A solution of 4.52 grams (0.0100 mole) of the maleic anhydride adduct of 3-beta-acetoxy-5,7,9(l1)-pregnatrien- 20-one, M. P. 263264.5 degrees centigrade, in a mixture of milliliters of l,4-dioxane and 400 milliliters of water containing four grams (0.10 mole) of sodium hydroxide was allowed to stand at room temperature for two and one-half hours, whereupon a small'quantity of plate-like crystals formed. These were dissolved by heating the mixture to seventy degrees centigrade for onehalf hour. The reaction mixture was then made acid with fifty milliliters of three normal hydrochloric acid and refrigerated to give a precipitate of 3.05 grams of needlelike crystals melting at 173-177 degrees centigrade. On crystallization from a dioXane-water mixture, the compound melted at 211-215 degrees centigrade. The melting point was found to vary somewhat with the rate of heating.

Analysis: Calculated for CztHazOt: 70.07; H, 7.53

Found: C, 69.80;.H, 7.47

A suspension of 0.400 gram of the maleic acid adduct of 3-beta-hydroxy-5,7,9(1l)-pregnatrien-20-one, in fifty milliliters of dry ether, was cooled in an ice-salt bath while a slight excess of diazomethane in methylene chloride was added over a 25-minute period with stirring. Ten minutes after addition was complete, the solution was placed on a steam bath and concentrated rapidly to dryness. The residue was crystallized from an acetonewater mixture to give 0.34 gram of the dimethyl maleate of 3-beta-hydroxy-5 ,7 ,9 1 1 )-pregnatrien-20-one, melting at 193195 degrees centigrade. After chromatography and recrystallization, the compound melted at 192-197 degrees centigrade.

In the same manner as given above, other dialkyl m'aleates, e. g., the diethyl, dipropyl, diisopropyl, dibutyl and dioctyl maleates of 3-hydroxy- 5,7,9(11)-pregnatrien-20- one are prepared from 3-hydroxy-5,7,9(ll)-pregna'trien- 20-one maleic acid adduct and the appropriate diazoalkane, or by other equivalent esterification procedure.

PREPARATION 15.DIMETHYL MALEATE 0F 3-BE-TA- acnroxr-S ,7 ,9 1 1 -PREGNAI'RIEN-20-ONE A solution of 0.15 gram of the dimethyl maleate adduct of 3-beta-hydroxy-5,7,9(11)-pregnatrien-20-one, in 2.5 milliliters of acetic anhydride and 2.5 milliliters of pyridine, was heated on the steam bath for ninety minutes, cooled to room temperature, and poured into ice-water. The resulting precipitate was collected by filtration and found to melt at 205-209 degrees centigrade. Recrystallization from methanol gave the dimethyl maleate of B-beta-acetoxy 5,7,9(11) IegnatriemZO-One,inelting at 207-211 degrees centigrade. Analysis: Calculated for C29H32O'7: C, 69.86-H,"7.68 Found: C, 69.81; H, 7.86

.water; dried, and' evaporated to' dryness.

11 By the'same manne'r of esterification, the-followingC-3 esters- 'were preparedt 1=)-dimethy1 maleate adductof 3- beta-forrnoxy'5 ,7 ,9(1:1 )-pregnatrien-20-one, meltingpoint 223230 degreeseen-tigrade; and'( 2) the dimethyl maleate adduct of 3-betabenzoyloxy-5 ;7,9 (-11 -pregnatrien-20-one, melting point 250 254 degrees centigrade.

PREr'ARATIoN 1,6.MALEIC ANHYDRIDE Aooucr or B-BETA- HEPTANoYLoxY-5;7,9( 1 1 PREGNATRIEN- 20-0Nr.

The maleic. anhydride; adduct of 3-beta-heptanoyloxy- 5,7,9(11.)-pregnatriena20=one, melting point 170-171 degrees centigrade,.was preparedzby refluxing the maleic acid adduct of 3-beta-hydroxy-5,7,9(1.1)-pregnatrien-20- one with heptylic anhydride and pyridine for a period of twenty hours, .and-working upthe reactionproduct in the usual'mannerv PREPARATION 1.7.--MALEIc ANHYDRIDE ADDUCT or 3-BarA- I'IYoRoxir 5',7,"9f( 1'1 -PREGNATRIEN-20-ONE Similarly; tlie maleic anhydride adduct of 3-beta-hydroxy,-5,7;'9(11i)?pregnatrien=20rone, melting point about 195 degrees centigrad'e; was.prepared by refluxing the maleicacid adduct of 3.-beta:hydroxy+5,7,9(11)-pregnatried-20mm with Dowthermtor eight hours; The 3-hydroXy-rnaleic anhydridetadduct is also obtained by heating -the- 3-hydroxy'maleie acid adduct' to just above its melting; po'int,,which: procedure. causes water to be' evolved, with the,closingofcthe anhydride ring.

In'the'samemanneras given above, still other 5,7,9- (11)-pregnatrien-20-'one adducts are prepared from the corresponding: 3;22;-diacyloxybisnor 5,7,9( 11 ,20 (22)- cholatetraene maleic acid, maleic anhydride and maleic acid diester adducts-s. Such compounds include the 3- f0rmoxy-5,7,9( 1 1 )-pregnatrien-20-one maleic acid; maleic acid. anhydride, dimethyl'maleate, diethyl maleate, dibutyl' maleate, dioctylim'aleate, diisopropyl maleate, and like adducts; the corresponding 3-propionoxy, butyroxy, valeroxy; hexanoyloxyg heptanoyloxy, octanoyloxy, naphthoyloxy; b'enzoyloxy, and similar 20-ketone'adducts, including, for example, 3-propionoxy-5,7,9 1 1 -pregnatrien- ZO-one dipropy-l maleate, 3-benzoyloxy-5,7,9(11)-pregna trien-2'0-one dibenzyl. maleate, S-heptan'oyloxy-S ,7 ,9( 1 1 pregnatrien-ZO-one dimethylmaleate, 3-valeroyloxy-5 ,7,9- (1'1) pregnatrien-ZO One. maleic acid anhydride adducts, and'the like.

Themaleic: anhydride: adduct of. 3-beta-acetoxy-5,7,9- (-1.1) -pregnatrien-2orone (0.45 gram, .001 mole) and 1.33 grams of lead tetraacetate were: added to 35 milliliters of 98.3 percent acetic acid which had been distilled over chromiumdrioxide. The mixture was heated in a water bath maintained. at: about 56 degrees centigrade andallowed tostand'at that temperature for seventy-two hours. The lead tetraacetate dissolved slowly during the first-hour and went: into solution. The solution was then evaporated-in vacuo until only'a few milliliters of solvent remained, the residue was dissolved in ether, washed with Crystallization of the residue from alcohol gave 0.17 gram of maleic anhydride adduct of 3-beta-,21-diacetoxy-5,7,9(11)-pregnatrien-2'0-one,,rnelting at225 to 235 degreescentigrade. After several recrystallization from'methanol, the material melted'at'240-2'43 degrees centigrade.

Analysis:

In:the mannergof the preceding'exarnple, the dimethyl maleate adduct of 3;-beta-acetoxy-5,7,9.(11)-pregnatrien- 20-one was convertetttoi thedimethiyl maleate adduct of 1 2 3-beta-,21-diacetoxy-5,7,9(11)'-pregnatrien-20-one, which melted at 210 -214 degrees Centigrade.

Analysis: Calculated for C31H40092' C, 66.89; H, 7.14

Found: C, 66.72; H, 7.24 C, 67.09; H, 7.34

In the same manner as given above, other 3-acyloxy, 21'-acetoxy-5,7,9(11)-pregnatrien-20-one adducts, including 3-formoxy,21-acetoxy 5,7,9(ll) pregnatrien-ZO-one maleic anhydride adduct,v 3-propionoxy,21-acetoxy- 5,7,9,(ll)-pregnatrien-20-One maleic acid adduct, 3- butyroxy,2l-acetoxy-5,7,9(11)-pregnatrien-20-one maleic anhydride adduct, 3-octanoyloxy,21-acetoxy-5,7,9(1'1)- pregnatrien-20'-one maleic anhydride adduct, 3-benzoyl- 0xy,21-acetoxy-5,7,9( 1 1 )-pregnatrien-20-one maleic anhydride adduct, and 3-valeroyloxy,21-acetoxy-5,7,9(ll)- pregnatrien-20-one maleic anhydride adduct, are prepared from the corresponding starting compound which is unsubstituted in the 21 position.

PREPARATION 20.-3- BETA ACETOXY-2l-HYDROXY-5,7 ,9-

(ll)-PREGNATRIEN-20-ONE MALEIC ACID ADDUCT One gram of 3-beta-21-diacetoxy-5,7,9(11)-pregnatrien-ZO-one maleic anhydride adduct was dissolved in milliliters of dioxane and the solution cooled to just above the freezing point of dioxane. A second solution consisting of 25 milliliters of 1.5 normal sodium hydroxide solution and 350 milliliters of water was cooled in an ice-bath and then added to the dioxane solution. The volume of the reaction mixture was then brought up to 500 milliliters by addition of water and allowed to stand at zero degrees Centigrade for one hour. The speed of the saponification is followed by removing aliquots at different time intervals and titrating with standard hydrochloric acid. At one hour, three equivalents of base had been used up. The reaction mixture was made acid to Congo red paper with ten percent hydrochloric acid and was concentrated on the steam bath in vacuo to about one hundred milliliters. This caused crystallization to take place, and, after cooling, 061 gram of 3-beta-acetoxy-2l-hydroxy-5,7,9(11)-pregnatrien-20-one maleic acid adduct, melting point 235240 degrees centigrade, was obtained.

PREPARATION 21.-3 BETA AcEToxY-21-IIYDRoxY-5,7,9-

(11)-PREGNATRIEN20-ONB MALEIC ANHYDRIDE ADDUCT Without further purification the acid thus produced in the preceding preparation was placed in a vacuum oven and heated for ten hours at 136140 degrees centigrade at eight to ten millimeters of pressure. The resulting product was recrystallized from acetone-water and gave 0.43 gram, melting point 225-233 degrees Centigrade, of 3-beta-acetoxy-2l-hydroxy 5 ,7,9(l1) pregnatrien 20 one maleic anhydride adduct. 'After two. recrystallizations from ethyl acetate-hexane, the product melted at 244-2 18- degrees centigrade. Infra-red analysis conclu+ sively showed that the 21-acetoxy group had been removed.

PREPARATION 22.3 -BETA-,2 1'-DIHYDROXY-5 ,7 ,9( 1 1 -PREG- NATRIEN-ZO-ONE MALEIC ACID AND ANHYDRIDE Aonucrs Example 1 .--3,21 -diacetoxy- ,7 ,9( I 1 )-pregnatrien-20-0ne (a) DIMETHYLANILINE A solution of'0.76 gram of the maleic anhydride adduct of 3,21-diacetoxy-5,7,9 (11)-pregnatrien-20-one in eight milliliters of dimethylaniline was heated under reflux for four and one-half hours under an atmosphere of nitrogen and the excess amine removed by distillation under reduced pressure. The residue was dissolved in 100 milliliters of ethyl ether, washed with equal volumes of cold one percent sulfuric acid, col-d five percent sodium carbonate solution, water, dried, and the solvent removed by distillation. Upon crystallization of the residue from fifteen milliliters of methanol, there was obtained 0.26 gram of 3,21 diacetoxy 5,7,9(1l)-pregnatrien-20-one, which melted at 134 to 138 degrees centigrade. When the product was recrystallized twice from methanol, its melting point was raised to 143.5 to 145.5 degrees centirade.

g ('b) N,N-DIMETHYLBENZYLAMINE A solution of one gram of the maleic anhydride adduct of 3,2l-diacetoxy-5,7,9(11)-pregnatrien-20-one in ten milliliters of N,N-dimethylbenzylamine was heated under reflux for four hours, and the excess amine removed under reduced pressure. The residue was dissolved in fifty milliliters of ethyl ether, washed with an equal volume of cold one percent sulfuric acid, cold five percent sodium carbonate solution, water, and dried. Upon removal of the solvent by distillation and crystallization of the residue from methanol, there was obtained 0.45 gram of 3,21-diacetoxy-5,7,9(11)-pregnatrien-20- one, which melted at 136 to 140 degrees centigrade,

In a like manner there can be obtained from the corresponding 3-acyloxy-21-acetoxy 5,7,9(1l) pregnatrien- 20-one maleic anhydride adduct, 3-formoxy-21-acetoxy- 5 ,7,9( 1 1 )-pregnatrien-20-one, 3-propionoxy-2l-acetoxy-S, 7,9(11) pregnatrien-20-one, 3-butyroxy-2l-acetoxy-5,7,9 (11) pregnatrien-ZO-one, 3-va1eroyloxy-21-acetoxy-5,7,9 l 1)-pregnatrien-20-one, 3-hexanoyloXy-21-acetoxy-5,7,9 1 1 )-pregnatrien-20-one, 3-heptano'yloxy-21 acetoxy-5,7,9 1 1 )-pregnatrien-20-one, 3-octanoyloxy-21-acetoxy-5,7,9 11 )-pregnatrien-20-one, 3-benzoyloxy-21-acetoxy-5,7,9 (11)-pregnatrien-20-one, and the like, as Well as the corresponding 3-acyloxy-21-hydroxy-5,7,9( 1 1)-pregnatrien- 20-ones, which are prepared by removal of the adduct radical from the corresponding maleic acid or maleic anhydride adducts of 3-acyloxy-21-hydroxy-5,7,9(11)-pregnatrien-ZO-ones, which are prepared as given for the 3- acetoxy-Zl-hydroxy adducts of Preparations 20 and 21.

Example 2.3,21-dihydroxy-5, 7,9 I I -pregnatrl'en-20-0ne The compound 3,21-diacetoxy-5,7,9( 1 1 -pregnatn'en- 20-one (0.618 gram) was dissolved in eighty milliliters of methanol and ten milliliters of four percent sodium hydroxide added thereto. After standing at room temperature for 1.5 hours, the solution was diluted with 135 milliliters of water and cooled. The compound 3,21-dihydroxy-5,7,9(11)-pregnatrien-20-one (0.340 gram, melting point 164-175 degrees centigrade) crystallized and was filtered from the solution. Several recrystallizations from dilute methanol or acetone-isopropyl ether raised the melting point to 175-185 degrees centigrade.

Analysis: Calculated for C21H2s03: C, 76.79; H, 8.59

Found: C, 76.82, 76.89; H, 8.55, 8.40

This compound is also prepared by removal of the adduct radical from the maleic acid or anhydride adduct of 3,21-dihydroxy-5,7,9 1 1)-pregnatrien-20-one, prepared as given above in Preparation 22, according to the method of the present invention.

In the same manner as given above in Example 2, 3-acetoxy-21-hydroxy 5,7,9(11)-pregnatrien-20-one can 14 be obtained from 3,21-diacetoxy-5,7,9(11)-pregnatrien- 20-one by carrying out the saponification under mild con: ditions with the use of only one molar equivalent of base, or as stated above after Example 1.

It is to be understood that the invention is not to be limited to the exact details of operation or exact compounds shown and described, as obvious modifications and equivalents will be apparent to one skilled in the art, and the invention is therefore to be limited only by the scope of the appended claims.

We claim:

1. A 3,2l-disubstituted-5,7,9(11)-pregnatrien-20-one of the formula:

5. Process for the production of a 3,2l-disubstituted- 5,7,9( 1l)-pregnatrien-20-one of the formula AcO wherein X is selected from the group consisting of hydropresence of an organic amine, at a temperature between about and 225 degrees centigrade, and separating the 3,21-disubstituted-5,7,9(11)-pregnatrien-20-one from the reaction product.

6. Process of claim 5, wherein the temperature is between about and 200 degrees centigrade.

7. Process of claim 5, wherein the amine is dimethylaniline.

8. Process of claim 5, wherein the starting adduct is a 3-acyloxy-21 -acetoxy-5 ,7 ,9 1 1)-pregnatn'en-20-one.

9. Process of claim 5, wherein the adduct is an adduct of 3 ,2 l-diacetoxy-S ,7 ,9 1 1)-pregnatrien-20-one.

10. Process of claim 5, wherein the adduct is an adduct of 3,21-dihydroxy-5,7,9 1 1)-pregnatrien-20-one.

No references cited. 

1. A 3,21-DISUBSTITUTED-5,7,9(11)-PREGNATRIEN-20-ONE OF THE FORMULA: 